Total RNA isolation from whole flies using TRIzol® (Life Technologies, Carlsbad, CA USA), cDNA synthesis, and real-time quantitative PCR were performed as described in Corl et al. [5]. RNA was isolated from 6–7 independent biological replicates of flies. Pre-designed 20× TaqMan® probe and primer mixes targeting Alk (Dm01797078_g1) and the ribosomal protein control transcript RpL32 (Dm02151827_g1) were purchased from Life Technologies. Data was normalized using the standard curve method and assessed for statistical significance using ANOVA.
